Separations of Active Compounds in Mulberry Leaves Using Centrifugal Partition Chromatography

• Main Authors: Wang, Bo-Sen, 王柏森
• Other Authors: Yu, Tiing
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/23pkkh

碩士 === 國立交通大學 === 應用化學系碩博士班 === 101 === Separations of active compounds, including Chlorogenic acid, Isoquercitrin, Astragalin, Quercetin 3-(6-malonylglucoside) and Kaempferol 3-(6-malonylglucoside) in mulberry leaves, were carried out using centrifugal partition chromatography (CPC) in this study. These five components in the crude extract by 60% (v/v) ethanol were first identified using HPLC-MS. After pre-extractions using ethyl acetate and water, we used solvent systems, (methyl t-butyl ether /acetone/H2O =6：4：10、7：3：10、8：2：10) and (methyl t-butyl ether /acetone/ H2O = 6：4：10、7：3：10、8：2：10) added with 0.6% (v/v) formic acid, four solvent systmes (methyl t-butyl ether /acetone/H2O/EA =7：3：10：2) , (methyl t-butyl ether /acetone/ H2O/EA = 7：3：10：2) added with 0.6% (v/v) formic acid, to separate sample extracts. The descending elution mode was applied, i.e. the lower aqueous layer was used as the mobile phase, and the upper organic layer as the stationary phase. Under 900 rpm, the volume of the stationary phase retained was 188 mL in the separation cells of a total volume 240 mL; stationary-phase retention ratio was 78%. The CPC effluent monitored using an on-line UV/Vis detector was fraction-collected every 3 mL, and the fractions were further analyzed using HPLC . The purities were 70.2%, 97.3%, 97.2%, 69.1% and 69.6% for Chlorogenic acid, Isoquercitrin, Astragalin, Quercetin 3-(6-malonylglucoside) and Kaempferol 3-(6-malonylglucoside), respectively, under the optimized separation.